JPH04178324A - Platelet coagulation-inhibitory agent - Google Patents

Abstract

PURPOSE:To provide the title agent combined with calcium antagonistic activity, containing, as active ingredient, a specific phenol derivative (or its nontoxic salt) such as eugenol, an active ingredient in rhizome of Asiasarum sieboldi being used as a crude drug. CONSTITUTION:The objective agent containing, as active ingredient, a phenol derivative (or its nontoxic salt) of formula I (R is H or methyl; R' is H, methyl, OH, methoxy, ethoxy or acetyl; n is 1-3), formula II or formula III (an example for the formula I: eugenol; an example for the formula II: 2-allyl-4-methylphenol; an example of the formula III: peonol). Of these active ingredients, eugenol, methyleugenol and peonol have also high-level calcium antagonistic activity other than platelet coagulation-inhibitory activity. The present agent is sufficiently highly active even at low dosage, being also suitable for patients with a combination of stenocardia, hypertension, cerebrovascular disease, etc.

Description

DETAILED DESCRIPTION OF THE INVENTION [Industrial Field of Application] The present invention is directed to a novel platelet aggregation inhibitor, and more particularly, to a platelet aggregation inhibitor that can also be used as a calcium antagonist because it has an effective calcium antagonistic effect. Regarding an aggregation inhibitor.

[Conventional technology] Conventionally, platelet prostaglandin (PG) inhibitors such as aspirin and flurbiprofen, ticlopidine and dipyridamole, etc. Platelet c A M
Drugs that increase P have been used as platelet aggregation inhibitors. Among these drugs, aspirin, which has a relatively high platelet aggregation inhibitory effect and is low in cost, is commonly used today.

Platelet aggregation inhibitors are used not only for small artery thrombosis in adults and Kawasaki disease in children, but also for hemolytic uremic syndrome, chronic renal glomerular disease, and after artificial valve or blood vessel augmentation surgery. Ta. However, aspirin, which has traditionally been commonly used as a platelet aggregation inhibitor, cannot be said to have sufficient platelet aggregation inhibitory activity for these diseases, and may have side effects such as liver dysfunction and bleeding tendency. The problem was that it was highly sensitive.

[Problems to be Solved by the Invention] The present invention solves the problems of the prior art described above, and aims to provide a platelet aggregation inhibitor that has high platelet aggregation inhibitory activity and is less likely to cause side effects when administered to humans. The purpose is

[Means for Solving the Problems] In order to solve the above-mentioned problems, the present inventors conducted intensive research and found that methyleugenol derivatives, which are the active ingredients of chili pepper, which has been used as an herbal medicine in Chinese medicine since ancient times, have been found to be effective against collagen in blood platelets. It was discovered that this compound has a remarkable inhibitory effect on aggregation (CPA) and an antagonistic effect on calcium, and was able to provide the present invention.

That is, the present invention provides a platelet aggregation inhibitor containing a phenol derivative represented by any of the following general formulas or a nontoxic salt thereof as an active ingredient, a platelet aggregation inhibitor that also has a calcium antagonistic effect, a calcium antagonist, and a platelet aggregation inhibitor. The present invention provides a calcium antagonist that also has an inhibitory effect.

(Space below) General formula: (However, in the formula, R represents a hydrogen atom or a methyl group, and R
' is a hydrogen atom, methyl group, hydroxy group, methoxy group,
1 selected from the group consisting of ethoxy group and acetyl group
Represents a species or two or more same or different groups,
n represents a natural number from 1 to 3. ) Suitable non-toxic salts of the compound that are effective in the platelet aggregation inhibitor of the present invention include alkali metal salts such as sodium salts and potassium salts, alkaline earth metal salts such as calcium salts, and the like.

The compound serving as the active ingredient of the platelet aggregation inhibitor of the present invention is
For example, eugenol, methyleugenol, 4-allylphenol and 4-allyl-2゜6-simethoxyphenol are represented by the general formula (1), and 2-allyl-4-
Methylphenol is represented by the general formula (n), and paeonol, catechol, 4-methoxyphenol, 2-methoxyphenol, etc. are represented by the general formula (III).

It has been recognized that all of the above compounds tend to have calcium antagonistic effects, and among these, eugenol, methyleugenol, and paeonol in particular have a high level of calcium antagonistic activity in addition to platelet aggregation inhibitory effects. confirmed.

The compound serving as the active ingredient of the platelet aggregation inhibitor of the present invention is
It can be easily obtained by organic synthesis or extraction from herbal medicines such as chili pepper, clove, and peony bark. However, in the present invention, the method for producing the compound or its nontoxic salt is not particularly limited as long as its purity and properties can be ensured.

The platelet aggregation inhibitor of the present invention can be prepared into an appropriate dosage form such as a tablet, gunpowder, powder, granule, capsule, liquid, or injection depending on the method of administration thereof. In addition, these drugs can be prepared by a conventional method using a compound represented by general formula (i), (II) or (m) as an active ingredient and a suitable carrier and/or excipient. can.

The dosage of platelet aggregation inhibitors or calcium antagonists prepared in various dosage forms varies depending on a wide variety of factors such as the type of active ingredient, the symptoms to be treated, age, sex, and body weight, but in general, ~1500mg/day
, (adult).

[Effect j] The platelet aggregation inhibitory activity of the compound serving as the active ingredient of the platelet aggregation inhibitor of the present invention is about several times as high as that of aspirin, a conventional platelet aggregation inhibitor. ing.

In addition, the calcium antagonistic activity of the compound serving as the active ingredient of the platelet aggregation inhibitor of the present invention is greater than that of Bapaverine, which has been conventionally used as a calcium antagonist.
It is equivalent to or slightly weaker than in, and even if this compound is contained as an active ingredient as a calcium antagonist, its medicinal efficacy is sufficiently effective. Therefore, the platelet aggregation inhibitor of the present invention that also has calcium antagonistic effects,
When administered to patients with arrhythmia, angina pectoris, hypertension, and cerebrovascular disorders, more favorable effects can be produced. Therefore, the platelet aggregation inhibitor can be used as a new drug for the circulatory system.

Furthermore, the compound serving as the active ingredient of the platelet aggregation inhibitor of the present invention is obtained by extraction from a compound that has long been used as a herbal medicine in Chinese medicine, and moreover, it has a high platelet aggregation inhibitory effect in a small amount. The possibility of side effects occurring when administered to humans is significantly reduced.

The compound serving as the active ingredient of the platelet aggregation inhibitor and calcium antagonist of the present invention can be produced as follows.

[Production Example 1] A method for producing methyleugenol will be described as an example of a method for producing methyleugenol by extracting a compound that is an active ingredient of the platelet aggregation inhibitor and calcium antagonist of the present invention.

First, 1 kg of chili was coarsely ground, and then extracted under reflux three times using about 5 times the amount of n-hexane as an extraction solvent.

Next, the solvent n-hexane was distilled off under reduced pressure from the n-hexane extract obtained by three times of □ extraction to obtain about 20 g of hexane extract.

The above hexane extract was fractionated by silica gel column chromatography, elemental analysis and IR spectrum were measured, and qualitative analysis was performed by comparing the results. First, n-hexan: EtOAc (5: l → 0:
10) EtOAc: MeOH (L:1 →0:1
The hexane extract was divided into 7 fractions using 0) as the eluent.

Next, the third fraction among these seven fractions was further subjected to silica gel column chromatography to obtain n-hexan:E
Three fractions were separated using tOAe (10:1) eluent.

Among the three fractions, the second fraction (200
0mg) elemental analysis and IR spectrum measurement,
When the results were compared with the standard product, it was confirmed that it was methyleugenol.

The above elemental analysis and IR spectrum measurement results are shown below.

Elemental analysis result Measured value...C・74.14%, H: s
, Theoretical value calculated as te%CIIH1402...
- C: 74.20%, H: 7.86% IR spectrum measurement result IR (CHCL3) 70m: 1150.1139.
1027 (Methoxygr), 917 (Al
lyl gr, ) [Production Example 2] Production of eugenol will be explained as another method by extracting a compound that is an active ingredient of the platelet aggregation inhibitor and calcium antagonist of the present invention.

First, 1 kg of cloves was subjected to reflux extraction three times with about three times the amount of n-hexane as an extraction solvent. Next, the n obtained by three extractions
The solvent n-hexane was distilled off from the -hexane extract under reduced pressure to obtain about 53 g of hexane extract.

The hexane extract was fractionated by silica gel column chromatography, subjected to elemental analysis and IR spectrum measurement, and qualitative analysis was performed by comparing the results. First, n-hexan: EtOAc (9.1-”l:
10) was used as the eluent, the hexane extract was divided into 5 fractions. Of these five fractions, the second fraction (approximately 35 g), which is an oily active substance, was subjected to thin layer chromatography and IR spectrum measurements, and the results were compared with the standard product, and it was confirmed to be eugenol. . The measurement results of thin layer chromatography and IR spectrum are shown below.

For thin layer chromatography, Kies is used on the plate.
Elgel 60 F294 (Merck) was developed for 10 cm using n-hexan:EtOAc (4:L) as a developing solvent, and a ferric chloride test solution was sprayed thereon as a coloring agent.

(Leaving space below) Thin layer chromatography result Rf value 0.32 and purplish-brown sample spot (consistent with eugenol) IR spectrum measurement result IR (Fi Im)/mark... ShiO-H: 3570 Shi C-C:1510 C-0・1265.1245.1035νC-H:9
12 [Production Example 3] As yet another production example of the method for extracting compounds that are active ingredients of the platelet aggregation inhibitor and calcium antagonist of the present invention, the production of paeonol will be described.

First, 1 kg of peony bark was roughly ground and then extracted under reflux three times with about three times the amount of 70% methanol as an extraction solvent. The methanol solvent was distilled off under reduced pressure from the methanol extract obtained in these three extractions, and the methanol extract was
I got g. Next, the methanol extract was partitioned between water and n-hexane, and each solvent was distilled off to obtain about 18 g of a hexane-transferred portion.

50% ethanol was added to this hexane transfer zone to crystallize it, and the active substance in the form of mp49-50m colorless needles was obtained by adding about 13
I got g. It was confirmed that this active substance was paeonol by mixing it with a standard product and comparing the results of thin layer chromatography and IR spectrum measurements.

The measurement results of the above IR spectrum are shown below.

I R (K B r )/cm...3400,29
40,1630,1570.1500 [Production Example 4] As an example of a method for organic synthesis of a compound serving as an active ingredient of the platelet aggregation inhibitor and calcium antagonist of the present invention, 4-
The production of allylphenol will be explained.

First, under anhydrous conditions, 20g of catechol was
Dissolve in methylene chloride from Step 1 and cool on ice. Add 50 g of boron tribromide little by little using a dropping funnel, and add HB.
When no r gas is released, stir at room temperature for 1 hour. After stirring, the mixture is concentrated under reduced pressure at 40°C to obtain crystals of 2-bromo-1,3,2-benzocyoxybrole. Dissolve the crystals in 180 ml of methylene chloride,
4.0 g of 4-allylanisole and 30 g of trifluoride diethyl ether complex were added to the mixture, and the mixture was stirred at room temperature for 48 hours. Next, this reaction solution is poured into a solution of hexane and water: 1=1, and the resulting organic layer is washed with water and physiological saline. After washing, this organic layer is extracted with an aqueous sodium hydroxide solution, further washed with hexane, and then neutralized with an aqueous hydrochloric acid solution while cooling with water. This neutralized solution was extracted with methylene chloride, and the extracted solution was dried over sodium sulfate and then concentrated under reduced pressure at 40°C to obtain 25 g of 4-allylphenol in the form of an oil.

Hereinafter, the present invention will be explained in more detail with reference to Examples. However, the scope of the present invention is not limited by the following examples.

[Example 1] As an example of the present invention, Table 1 shows the method and results of a platelet aggregation inhibitory activity test of the active ingredient compound of the platelet aggregation inhibitor of the present invention.

In this example, a platelet aggregation inhibitory activity test was conducted using the following method. Centrifugation of fresh rabbit whole blood (1200 rpm, 10
The anti-aggregation activity of the compounds shown in Table 1 was evaluated in vitro using platelet-rich plasma (PRP) obtained by M1n) as a platelet sample. In vitro evaluation of antiaggregation activity was performed using platelet-rich plasma A
By nephelometry of DP aggregation and Kolaken aggregation, Pay
ton Aggregation Module (M
ode1300B) was used as a measuring instrument. In addition, 1 ml of agglutination reaction mixture contains 10 μg of platelet-rich plasma,
Soluble collagen 10■ or 0.2A ADP, 98
Test substance dissolved in 0.0% ethanol or DMSO.
It contained 5 tlM. Furthermore, in this test, it was determined that there was aggregation inhibitory activity when the aggregation inhibition rate exceeded 50%, and aspirin, which has been conventionally used as a platelet aggregation inhibitor, was selected as a positive target substance.

Table 1 shows the results of the platelet aggregation inhibitory activity test conducted by the above method.

(The following is a blank space) Table 1 As can be seen from Table 1, the substances measured in this example were confirmed to exhibit significantly higher platelet aggregation inhibitory activity than aspirin. More specifically, the I C5o (M) value of the platelet aggregation inhibitory effect of eugenol showed approximately 18 times the activity of aspirin, and also showed sufficient activity of 4-allylphenol, about 7 times.

[Example 2] As another example of the present invention, the method and results of a calcium antagonist activity test of the active ingredient compound of the platelet aggregation inhibitor of the present invention are shown using Table 2.

In this example, the removed caecum 1fJ (taenj) of a guinea pig was used.
A calcium antagonist activity test was carried out by the Magnus method using a suspended strain of C. acoli. First, the cecum fI (taenia coli) was removed from a male guinea pig weighing 300 g, and the Ca"-free KCI-Locke-Ri
nger solution (KCI 159.6 MgCh2, I
Na1(Co, 5.9 glucose 2.8mM
) and kept at 30° C. for about 60 minutes to stabilize while aerating a 95% oxygen-5% carbon dioxide gas mixture.

After stabilization, add 0.3 m of CaCl□ solution (0.1 g/ml)
1 was added and the shrinkage that occurred was recorded and designated as A.

Next, do this three times or more every 10 minutes.Ca”-rree KC
Wash with I-Locke-Ringer solution, and when stable, add test sample solution. Same <CaCl2 after 3 minutes
Add 0.3 ml of the solution and record the contraction that occurs,
This was designated as B. A and B thus measured were substituted into the following formula to calculate the inhibition rate, and the results are shown in Table 2. Note that Papaverin, which has been conventionally used as a calcium antagonist, was used as a positive target substance.

a formula.

(A-B/A)X100 (%) - Inhibition rate (1%)
Table 2 From Table 2 above, it was confirmed that the calcium antagonistic effect of the compound of this example was slightly weaker than that of Papaverin, and that its medicinal efficacy was sufficiently effective.

[Effects of the Invention] The platelet aggregation inhibitor of the present invention has an extremely high activity of inhibiting platelet aggregation due to the compound serving as its active ingredient, so platelet aggregation can be sufficiently inhibited even in small amounts. Ta. In addition, some of the compounds that are effective in the platelet aggregation inhibitor of the present invention also have calcium antagonistic activity, and their calcium antagonistic activity is sufficiently effective as a calcium antagonist whose medicinal effect is only calcium antagonistic activity. Therefore, it can have a favorable effect even on patients who also have diseases such as angina pectoris, hypertension, and cerebrovascular disorders. Furthermore, the compound serving as the active ingredient of the platelet aggregation inhibitor of the present invention can be organically synthesized or extracted from herbal medicines such as chili pepper, clove, and peony bark.
Stable products can be obtained at low cost using simple means.

Claims (4)

[Claims] (1) The following general formulas: ▲There are mathematical formulas, chemical formulas, tables, etc.▼……(I) or ▲There are mathematical formulas, chemical formulas, tables, etc.▼……(II) or ▲There are mathematical formulas, chemical formulas, tables, etc.▼ ...(III) (However, in the formula, R represents a hydrogen atom or a methyl group, and R
' is a hydrogen atom, methyl group, hydroxy group, methoxy group,
1 selected from the group consisting of ethoxy group and acetyl group
Represents a species or two or more same or different groups,
n represents a natural number from 1 to 3. ) A platelet aggregation inhibitor comprising a phenol derivative or a non-toxic salt thereof as an active ingredient. (2) The platelet aggregation inhibitor according to claim 1, which also has a calcium antagonistic effect. (3) The following general formulas: ▲There are mathematical formulas, chemical formulas, tables, etc.▼……(I) or ▲There are mathematical formulas, chemical formulas, tables, etc.▼……(II) or ▲There are mathematical formulas, chemical formulas, tables, etc.▼ ...(III) (However, in the formula, R represents a hydrogen atom or a methyl group, and R
' is a hydrogen atom, methyl group, hydroxy group, methoxy group,
1 selected from the group consisting of ethoxy group and acetyl group
Represents a species or two or more same or different groups,
A calcium antagonist comprising a phenol derivative or a non-toxic salt thereof as an active ingredient. (4) The calcium antagonist according to claim 3, which also has a platelet aggregation inhibiting effect.